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EXPRESSION OF RECOMBINANT HUMAN ERYTHROPOIETIN WITH GLYCOSYLATION MODIFICATION IN HEK293T CELLS
Get PDFStability of erythropoietin (EPO) depends on its glycosylation states. With more glycosylation sites, the EPO protein will be more stable and also increase its half-life. A construct of recombinant human erythropoietin (rhEPO) which contains 2 additional N-link for glycosylation were designed. Based on translation analysis using ORF (open reading frame)-finder and protein alignment analysis using blast-p of NCBI home page, expected recombinant hEPO with additional 6-histidin tag in carboxyl terminus was expressed. HEK293T cells were transfected with recombinant plasmid containing rhEPO by using calcium phosphate method. Expression of rhEPO was detected by dot blot and Western blot analysis using hEPO antibody as the primary antibody and antirabbit antibody with alkaline phospatase linked as the secondary antibody. The bands were detected by BCIP/NBT color development substrate. The data indicated detection of EPO in culture medium of transfected HEK293T cells.Key words: HEK293T cell, calcium phosphate transfection, N-linked glycosylation, recombinant human erythropoieti
APPLICATION OF FOURIER TRANSFORM INFRARED SPECTROSCOPY FOR QUALITY CONTROL OF PHARMACEUTICAL PRODUCTS: A REVIEW
Get PDFAnalysis of pharmaceutical products covers all aspects of quality control of active pharmaceutical ingredients (API) and finished products. Today, Fourier transform infrared (FTIR)spectroscopy, especially in combination with chemometrics software, has emerged as one of the romising analytical techniques to be used in pharmaceutical industry, for quality control of desired pharmaceutical products. Compared with other instrumental techniques, FTIR spectroscopy offers some advantages, namely it is rapid, simple in sample preparation, and not destructive. In this review, the application of FTIR spectroscopy for qualitative and quantitative determinations of API and monitoring drug release are described.Key words: FTIR spectroscopy, quality control, pharmaceutical products
DEVELOPMENT AND EVALUATION OF IN SITU GELS OF MOXIFLOXACIN FOR THE TREATMENT OF PERIODONTITIS
Get PDFGel dosage forms are successfully used as drug delivery systems to control drug release and protect the medicaments from a hostile environment. The main objective of the present investigation is to formulate and evaluate in situ gels of moxifloxacin for the treatment of periodontitis using gellan gum and sodium alginate based on the concept of ion activated systems. The system utilizes polymers that exhibit sol-to-gel phase transition due to change in specific physico-chemical parameters. Sol-to-gel transformation occurred in the presence of monovalent/divalent cations. It was found that increase in the concentration of calcium ions produced stronger gels. Formulations were evaluated for gelling capacity, drug content, clarity, viscosity, gel strength, spread ability, microbiological studies and in vitro release. The results shown by the above characterization studies were found to be satisfactory. Experimental part showed that viscosity of sols and gel strength was increased with increase in the concentration of polymers and the sustained release of drug was observed. The formulations were therapeutically efficacious, sterile and provided sustained release of the drug over a period of time. These results demonstrated that the developed system is an alternative to conventional drug delivery systems, provides patient compliance and economical.Key words: in situ gels, moxifloxacin, periodontitis, polymers, Ion activation
BIOCHEMICAL CHANGES OF CERTAIN PLANT SPECIES DUE TO VARIOUS DRYING TREATMENTS
Get PDFPhytochemical analysis of the Indian medicinal plants to identify the therapeutically active chemical compounds in them and clinically testing for their efficacy and risks is an inescapable procedure, to provide new sources of natural products to the pharmaceutical industry. Therefore, the phytochemical screening studies should be concentrated on the hitherto unexplored medicinal plants for their medicinal values. In the current study the leaves of different medicinal plants like Achyranthus aspera, Acalypha indica, Euphorbia hirta, Lindenbergia indica, Parthenium hysterophorus and Pesistrophe bicalyculata were taken for the chlorophyll and carotenoid estimation by the method of Jayaraman (1981). The leaves of the following plants were allowed to dry in an oven at different temperatures, time period and freeze drying. Oven drying was done at 50±10C for 9 hours and at 70±1ºC for 5 hours. Results obtained revealed that both the drying treatments affect/decrease the chlorophyll content of all the six plants. The maximum effect was seen in Euphorbia hirta (93.97%) for oven drying at 50±1ºC for 9 hours. The minimum effect was observed in Parthenium hysterophorus (21.68%) for oven drying at 70±1ºC for 5 hours and Achyranthus aspera (11.56%) for freeze drying. The leaf preparations treated by oven drying was found to have lower chlorophyll and carotenoid content as compared to freeze drying which in turn has lower chlorophyll content as compared to fresh extracts.Key words: Chlorophyll content, Drying treatments, and leaf extract.
FORMULATION OF NANOCURCUMIN USING LOW VISCOSITY CHITOSAN POLYMER AND ITS CELLULAR UPTAKE STUDY INTO T47D CELLS
Get PDFUsing of curcumin as anti cancer agent is restricted by its low solubility, therefore it has low bioavability. This obstacle can be solved by the development of curcumin nanoparticle. Nanoparticle technology has been started to be developed as an alternative solution to improve drug delivery pofile, especially for the less bio-available chemical. This study was aimed to develope nanocurcumin formulation with low viscosity chitosan as the matrix and to study its ability to be taken into the cells in vitro. Method used in the formulation of nanocurcumin in this study is by ionic gelation followed by freeze drying. Entrapment Efficiency then assayed, and its stability was tested by incubating the formula into artificial intestinal fluid (AIF). Furthermore, its toxicity was evaluated, also its cellular uptake ability into T47D cell line. It was found that the Entrapment Efficiency in acetate buffer at pH 4 is higher than at pH 5. This formula also has a good stability in AIF. For the cellular uptake study through fluorescence microscope, it was found that the complex has an ability to penetrate cellular membrane into the cytosol. The cytotoxicity study tell us that the nanocurcumin is non-toxic to normal cell line. For the characterization of the nanoparticles, the average size of this particle is 269.8 nm, its zeta-potential is +18.63 mV, with spherical particle morphology. From the result ofthis study, it is concluded that formulation of nanocurcumin using low viscosity chitosan polymer as the matrix has a great potential as an alternative for anticancer therapy.Key words: nanoparticle, curcumin, low viscosity chitosan, T47D cell line.
IDENTIFICATION OF SAFETY ALERT BY MONITORING ANALYTICAL PARAMETERS AND HIGH-RISK DRUGS
Get PDFEarly detection of adverse drug reactions (ADR) increases patient safety. Our objective was to identify ADR by monitoring laboratory parameters and high-risk drugs. We carried out a two-month prospective observational study in a Internal Medicine Department, with daily recording of drugs prescribed and the following parameters: Na, K, Ca, serum creatinine, glomerular filtration rate (GFR), INR, glucose, haemoglobin, platelets, ALT, AST, bilirubin, GGT, alkaline phosphatase, TSH, T4, and blood digoxin. High-risk drugs were closely monitored. 52 patients included, of whom 46.2% experienced an ADR. We observed an association with drugs in 25.5%, as follows: reduction in GFR, 26.9% (associated with loop diuretics [41.7%], angiotensin-converting enzyme [ACE] inhibitors [33.3%], angiotensin II receptor blockers [ARB] [16.6%], andanti-diabetic drugs [8.3%]); hypokalemia, 22.3% (associated with loop diuretics [50.0%], potassium-free fluid [37.5%], and salbutamol [12.5%]); hyperkalemia, 14.4% (associated with ACE inhibitors [60.0%] and ARB [40.0%]); INR out of range, 10.8% (associated with drug interactions [66.7%]); hyperglycemia, 8.1% (associated with corticosteroids [66.7%] and anti-diabetic drugs [33.3%]); and other conditions, 18.8%. We conclued that patient safety could be improved by implementing warnings in electronic prescriptions in cases of a decrease in GFR or modification of potassium levels in patients who are prescribed loop diuretics, ACE inhibitors, or ARBs.Key words: Adverse drug reaction, clinical decision support, high-risk drug, safety
DIFFERENTIATION BETWEEN PORCINE AND BOVINE GELATIN IN CAPSULE SHELLS BASED ON AMINO ACID PROFILES AND PRINCIPAL COMPONENT ANALYSIS
Get PDFGelatin used for capsule shells can be derived from porcine origin. Some religion like Islam prohibited their followers to consume any products containing porcine derivatives including porcine gelatin in the products. Consequently, some rapid and reliable techniques are continuously developed for detection of porcine gelatin in some pharmaceutical products like capsule shells. The aim of this study was to evaluate the possibility to use amino acid profile in combination with chemometrics of principal component analysis for detection and classification between gelatin in capsule derived from porcine and bovine. Based on score plot of first principal components (PC1) and second principal components (PC2), porcine and bovine gelatins in capsule could be apparently distinguished.Key words: bovine gelatin; porcine gelatin; amino acid profile; principal component analysi
THE OPTIMISED CONDITIONS OF INDUCTION OF RECOMBINANT RIP rMJC15310 ACTIVITY ISOLATED FROM Mirabilis jalapa L. LEAVES
Get PDFRibosome Inactivating Proteins (RIPs) are compounds isolated from plants with ability to inhibit protein synthesis. The inhibition of protein synthesis is due to inactivation of ribosomal RNA through a site-specific deadenylation mediated by RNA N-glycosidase. Reportedly, RIPs mainly possess wide range of bioactivity including antiviral activity against plant infections. Other activities of RIP were as abortifacien, antivirus and anticancer. This study was aimed to isolate and characterize the optimum conditions for inducing the expression of recombinant RIPs isolated from the leaves of Mirabilis Jalapa L. We have been successfully isolated several RIPs and engineered these proteins to be expressed in E. coli. These recombinant proteins were obtained by screening cDNA library originated from the mRNA of Mirabilis jalapa L leaves, and inserted into pUC19 carrying lacZ gene. The presence of recombinant plasmid was tested by using α-complementation assay. Many RIPs have been isolated from plants and these proteins express enzymatic activity by cutting supercoiled double stranded DNA. One RIP namely rMJC15310 was obtained from this study and the proteins having ~ 8kb in size, cut the supercoiled DNA into linear form at the concentration as low as 5 µg. The ability to cut supercoiled DNA increased on inducing its expression with 0.4% IPTG.Key words: Ribosome Inactivating Proteins (RIP), IPTG, Mirabilis jalapa L., recombinant protein
CHROMOSOME CHARACTERIZATION OF THREE VARIETIES OF GINGER (Zingiber officinaleRosc.)
Get PDFGiant ginger (Zingiber officinale Rosc. var. officinale), red ginger (Zingiber officinale Rosc. var. rubra) and small ginger (Zingiber officinale Rosc. var. amarum) are three varieties of Zingiber officinale Rosc. They have a lot of benefit and often used by Indonesian as a traditional drug. Moreover, they have a big chance to be use as a flavor in world wide. Therefore, research for their quality, quantity and continuity of supplyare needed. Characterization of their chromosomes is one effort for improving ginger cultivation. The objective of this research was to study mitotic time and chromosome characters of three varieties of ginger. Squashing method was used for chromosome preparation. The results showed that mitotic time of giant ginger is 09.00-10.05 am, red ginger is 09.00-10.30 am, while small ginger is 08.45-11.00 am. Chromosome number of giant ginger and small ginger are 2n=2x=30, while red ginger is 2n=2x=22. Giant ginger has R= 3,109, Red ginger has R = 3,206 and small ginger has R = 4,065. Based on chromosome characters it is revealed that relationship between giant ginger and red ginger is closer that of compare to small ginger. This result is important as basic information for improving the gingers production through breeding program.Key words: Zingiber officinale Rosc., mitotic time, chromosome characterization, squashmetho
PHYTOCHEMICAL STUDIES OF Acacia catechu
No full textAcacia catechu Willd. belongs to the family Mimosaceae. It is well known for the powder of wood known as Kathha” in India, which has wide applications. In the present work, the dry wood powder is extracted in various solvents such as 1,4-dioxane, methanol, dimethyl formamide (DMF), acetone, ethanol and distilled water. The phytochemical and physicochemical properties of these various solvent extracts were studied. The phyto constituents present in the different extracts were saponins, tannins, sugars, terpenes, catechu-tannic acid. Alkaloids were present in organic solvents only. The antibacterial activity of all the extracts in DMSO was studied against five bacterial strains but none of the extracts inhibit any of the studied bacterial strains.Key words: Acacia catechu, 1,4-dioxane, methanol, dimethyl formamide (DMF), acetone, ethanol, distilled water, phytochemical analysis, physicochemical analysis